Effect of proton beam on blood vessel formation in early developing zebrafish (Danio rerio) embryos

  • Gun Hyuk Jang
  • Ji-Hong Ha
  • Tae-Lin Huh
  • You Mie Lee
Articles Drug Efficacy and Safety
First Online:
Received:
Revised:
Accepted:

Abstract

Proton beam therapy can kill tumor cells while saving normal cells because of its specific energy delivery properties and so is used to various tumor patients. However, the effect of proton beam on angiogenesis in the development of blood vessels has not been determined. Here we used the zebrafish model to determine in vivo whether proton beam inhibits angiogenesis. Flk-1-GFP transgenic embryos irradiated with protons (35 MeV, spread out Bragg peak, SOBP) demonstrated a marked inhibition of embryonic growth and an altered fluorescent blood vessel development in the trunk region. When cells were stained with acridine orange to evaluate DNA damage, the number of green fluorescent cell death spots was increased in trunk regions of irradiated embryos compared to non-irradiated control embryos. Proton beam also significantly increased the cell death rate in human umbilical vein endothelial cells (HUVEC), but pretreatment with N-acetyl cystein (NAC), an antioxidant, reduced the proton-induced cell death rate (p<0.01). Moreover, pretreatment with NAC abrogated the inhibition of trunk vessel development and prevented the trunk malformation caused by proton irradiation. In conclusion, proton irradiation significantly inhibited in vivo vascular development possibly due to increased vascular cell death via reactive oxygen species formation.

Key words

Proton beam SOBP Blood vessel formation Cell death Reactive oxygen species 
This is a preview of subscription content, log in to check access.

References

  1. Belli, M., Bettega, D., Calzolari, P., Cera, F., Cherubini, R., Dalla Vecchia, M., Durante, M., Favaretto, S., Gialanella, G., Grossi, G., Marchesini, R., Moschini, G., Piazzola, A., Poli, G., Pugliese, M., Sapora, O., Scampoli, P., Simone, G., Sorrentino, E., Tabocchini, M. A., Tallone, L., and Tiveron, P., Inactivation of human normal and tumour cells irradiated with low energy protons. Int. J. Radiat. Biol., 76, 831–839 (2000).PubMedCrossRefGoogle Scholar
  2. Bettega, D., Calzolari, P., Chauvel, P., Courdi, A., Herault, J., Iborra, N., Marchesini, R., Massariello, P., Poli, G. L., and Tallone, L., Radiobiological studies on the 65 MeV therapeutic proton beam at Nice using human tumour cells. Int. J. Radiat. Biol., 76, 1297–303 (2000).PubMedCrossRefGoogle Scholar
  3. Brunner, T. B., Gupta, A. K., Shi, Y., Hahn, S. M., Muschel, R. J., McKenna, W. G., and Bernhard, E. J., Farnesyltransferase inhibitors as radiation sensitizers. Int. J. Radiat. Biol., 79, 569–576 (2003).PubMedCrossRefGoogle Scholar
  4. Brunner, T. B., Hahn, S. M., McKenna, W. G., and Bernhard, E. J., Radiation sensitization by inhibition of activated Ras. Strahlenther Onkol., 180, 731–740 (2004).PubMedCrossRefGoogle Scholar
  5. Cerimele, F., Battle, T., Lynch, R., Frank, D. A., Murad, E., Cohen, C., Macaron, N., Sixbey, J., Smith, K., Watnick, R. S., Eliopoulos, A., Shehata, B., and Arbiser, J. L., Reactive oxygen signaling and MAPK activation distinguish Epstein-Barr Virus (EBV)-positive versus EBV-negative Burkitt’s lymphoma. Proc. Natl. Acad. Sci. U.S.A., 102, 175–179 (2005).PubMedCrossRefGoogle Scholar
  6. Chadderton, N., Cowen, R. L., Sheppard, F. C., Robinson, S., Greco, O., Scott, S. D., Stratford, I. J., Patterson, A. V., and Williams, K. J., Dual responsive promoters to target therapeutic gene expression to radiation-resistant hypoxic tumor cells. Int. J. Radiat. Oncol. Biol Phys,. 62, 213–222 (2005).PubMedCrossRefGoogle Scholar
  7. Chan, J., Bayliss, P. E., Wood, J. M., and Roberts, T. M., Dissection of angiogenic signaling in zebrafish using a chemical genetic approach. Cancer Cell., 1, 257–267 (2002).PubMedCrossRefGoogle Scholar
  8. Concin, N., Stimpfl, M., Zeillinger, C., Wolff, U., Hefler, L., Sedlak, J., Leodolter, S., and Zeillinger, R., Role of p53 in G2/ M cell cycle arrest and apoptosis in response to gamma-irradiation in ovarian carcinoma cell lines. Int J Oncol., 22, 51–57 (2003).PubMedGoogle Scholar
  9. Di Pietro, C., Piro, S., Tabbi, G., Ragusa, M., Di Pietro, V., Zimmitti, V., Cuda, F., Anello, M., Consoli, U., Salinaro, E. T., Caruso, M., Vancheri, C., Crimi N., Sabini, M.G., Cirrone, G. A., Raffaele, L., Privitera, G., Pulvirenti, A., Giugno, R., Ferro, A., Cuttone, G., Lo Nigro, S., Purrello, R., Purrello, F., and Purrello, M., Cellular and molecular effects of protons: apoptosis induction and potential implications for cancer therapy. Apoptosis, 11, 57–66 (2006).PubMedCrossRefGoogle Scholar
  10. el Gamoussi, R., Stratford, I. J., and Guichard, M., Relationship between intracellular concentration and radiosensitizing effect of pimonidazole and etanidazole on two human melanoma cell lines. Int J Radiat Biol., 63, 27–36 (1993).PubMedCrossRefGoogle Scholar
  11. Engel, R.H. and Evens, A.M., Oxidative stress and apoptosis: a new treatment paradigm in cancer. Front Biosci., 11, 300–312 (2006).PubMedCrossRefGoogle Scholar
  12. Folkman, J., Angiogenesis and its inhibitors. Important Adv Oncol., 42–62 (1985).Google Scholar
  13. Giuliano, M., Lauricella, M., Vassallo, E., Carabillo, M., Vento, R., and Tesoriere, G., Induction of apoptosis in human retinoblastoma cells by topoisomerase inhibitors. Invest Ophthalmol Vis Sci., 39, 1300–1311 (1998).PubMedGoogle Scholar
  14. Haffter, P., Granato, M., Brand, M., and Mullins, M. C., The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio. Development., 123, 1–36 (1996).PubMedGoogle Scholar
  15. Harris, A. L., Hypoxia-a key regulatory factor in tumour growth. Nat Rev Cancer, 2, 38–47 (2002).PubMedCrossRefGoogle Scholar
  16. Hashimoto, T., Tokuuye, K., Fukumitsu, N., Igaki, H., Hata, M., Kagei, K., Sugahara, S., Ohara, K., Matsuzaki, Y., and Akine, Y., Repeated proton beam therapy for hepatocellular carcinoma. Int. J. Radiat. Oncol. Biol. Phys., 65, 196–202 (2006).PubMedGoogle Scholar
  17. Isenberg, J. S., Jia, Y., Field, L., Ridnour, L. A., Sparatore, A., Del Soldato, P., Sowers, A. L., Yeh, G. C., Moody, T. W., Wink, D. A., Ramchandran R., and Roberts D. D., Modulation of angiogenesis by dithiolethione-modified NSAIDs and valproic acid. Br J Pharmacol., 151, 63–72 (2007).PubMedCrossRefGoogle Scholar
  18. Jabs, T., Reactive oxygen intermediates as mediators of programmed cell death in plants and animals. Biochem Pharmacol., 57, 231–245 (1999).PubMedCrossRefGoogle Scholar
  19. Kari, G., Rodeck, U., and Dicker, A.P., Zebrafish: an emerging model system for human disease and drug discovery. Clin Pharmacol Ther., 82, 70–80 (2007).PubMedCrossRefGoogle Scholar
  20. Kennedy, A. R., Guan, J., and Ware, J. H. Countermeasures against space radiation induced oxidative stress in mice. Radiat Environ Biophys., 46, 201–203 (2007).PubMedCrossRefGoogle Scholar
  21. Kimmel, C. B., Ballard, W. W., Kimmel, S. R., Ullmann, B., and Shilling, T. F., Stages of embryonic development of the zebrafish. Devopmental. Dyn., 203, 253–310 (1995).Google Scholar
  22. Langheinrich, U., Zebrafish: a new model on the pharmaceutical catwalk. Bioessays., 25, 904–912 (2003).PubMedCrossRefGoogle Scholar
  23. Lee, K. B., Lee, J. S., Park, J. S., Huh, T. L., and Lee, Y. M., Low energy proton beam induces tumor cell apoptosis through reactive oxygen species and activation of caspases. Exp. Mol. Med., 40, 118–129 (2008).PubMedCrossRefGoogle Scholar
  24. Levin, W. P., Kooy H., Loeffler J. S., and DeLaney T. F., Proton beam therapy. Br.J.Cancer 93, 849–854 (2005).PubMedCrossRefGoogle Scholar
  25. Li, L. and Dowling, J. E., A dominant form of inherited retinal degeneration caused by a non-photoreceptor cell-specific mutation. Proc. Natl. Acad. Sci. U.S.A., 94, 11645–11650 (1997).PubMedCrossRefGoogle Scholar
  26. Limoli, C. L., Giedzinski, E., Baure, J., Rola, R., and Fike, J. R., Redox changes induced in hippocampal precursor cells by heavy ion irradiation. Radiat Environ Biophys, 46, 167–172 (2007).PubMedCrossRefGoogle Scholar
  27. Mishra, K. P., Cell membrane oxidative damage induced by gamma-radiation and apoptotic sensitivity. J Environ Pathol Toxicol Oncol. 23, 61–66 (2004).PubMedCrossRefGoogle Scholar
  28. Sanchez-Prieto, R., Rojas, J. M., Taya, Y., and Gutkind, J. S., A role for the p38 mitogen-acitvated protein kinase pathway in the transcriptional activation of p53 on genotoxic stress by chemotherapeutic agents. Cancer Res., 60, 2464–2472 (2000).PubMedGoogle Scholar
  29. Seiler, C. and Nicolson, T., Defective calmodulin-dependent rapid apical endocytosis in zebrafish sensory hair cell mutants. J Neurobiol., 41, 424–434 (1999).PubMedCrossRefGoogle Scholar
  30. Semenza, G. L., Intratumoral hypoxia, radiation resistance, and HIF-1α. Cancer Cell. 5, 405–406 (2004).PubMedCrossRefGoogle Scholar
  31. Taraboletti, G. and Giavazzi, R., Modelling approaches for angiogenesis. Eur. J. Cancer., 40, 881–889 (2004).PubMedCrossRefGoogle Scholar
  32. Wan, X. S., Zhou, Z., Ware, J. H., and Kennedy, A. R., Standardization of a fluorometric assay for measuring oxidative stress in irradiated cells. Radiat Res., 163, 232–240 (2005).PubMedCrossRefGoogle Scholar
  33. Westerfield, M., The zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio). (3rd ed.), University of Oregon Press, Eugene (1995).Google Scholar
  34. Williams, K. J., Telfer, B. A., Xenaki, D., Sheridan, M. R., Desbaillets, I., Peters, H. J., Honess, D., Harris, A. L., Dachs, G. U., van der Kogel, A., and Stratford, I. J., Enhanced response to radiotherapy in tumours deficient in the function of hypoxia-inducible factor-1. Radiother Oncol., 75, 89–98 (2005).PubMedCrossRefGoogle Scholar
  35. Yabu, T., Tomimoto, H., Taguchi, Y., Yamaoka, S., Igarashi, Y., and Okazaki, T., Thalidomide-induced antiangiogenic action is mediated by ceramide through depletion of VEGF receptors, and is antagonized by sphingosine-1-phosphate. Blood, 106, 125–134 (2005).PubMedCrossRefGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2008

Authors and Affiliations

  • Gun Hyuk Jang
    • 1
  • Ji-Hong Ha
    • 1
  • Tae-Lin Huh
    • 1
  • You Mie Lee
    • 1
  1. 1.School of Life Sciences and Biotechnology, College of Natural SciencesKyungpook National UniversityDaeguKorea

Personalised recommendations